Real-time in vivo green fluorescent protein imaging of a murine leishmaniasis model as a new tool for Leishmania vaccine and drug discovery.
Leishmania species are obligate intracellular protozoan parasites that cause a broad spectrum of clinical diseases in mammalian hosts. The most frequently used approach to quantify parasites in murine model systems is based on thickness measurements of the footpad or ear after experimental infection. To overcome the limitations of this method, we used a Leishmania mutant episomally transfected with enhanced green fluorescent protein, enabling in vivo real-time whole-body fluorescence imaging, to follow the progression of Leishmania infection in parasitized tissues. Fluorescence correlated with the number of Leishmania parasites in the tissue and demonstrated the real-time efficacy of a therapeutic vaccine. This approach provides several substantial advantages over currently available animal model systems for the in vivo study of immunopathogenesis, prevention, and therapy of leishmaniasis. These include improvements in sensitivity and the ability to acquire real-time data on progression and spread of the infection.
Mehta
Sanjay R
SR
Department of Medicine, University of California, San Diego, San Diego, CA, USA.
Huang
Robert
R
Yang
Meng
M
Zhang
Xing-Quan
XQ
Kolli
Bala
B
Chang
Kwang-Poo
KP
Hoffman
Robert M
RM
Goto
Yasuyuki
Y
Badaro
Roberto
R
Schooley
Robert T
RT
D000818:Animals; D004195:Disease Models, Animal; D055808:Drug Discovery; D005260:Female; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007891:Leishmania / Q000235:genetics / Q000302:isolation & purification*; D007896:Leishmaniasis / Q000276:immunology* / Q000469:parasitology*; D054332:Leishmaniasis Vaccines / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence / Q000379:methods*
D054332:Leishmaniasis Vaccines; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins